Yang Fu, Wang Fang, Guo Yingjun, Zhou Qi, Wang Yue, Yin Yixuan, Sun Shuhan
Department of Medical Genetics, Second Military Medical University, 800 Xiang Yin Road, Shanghai 200433, PR China.
J Proteome Res. 2008 Nov;7(11):4898-903. doi: 10.1021/pr800547v. Epub 2008 Oct 9.
Many recent studies have indicated that virus-like particles (VLPs) have many potential applications in the fields of vaccine development and gene therapy. However, we still know little about the subtle mechanisms involved in the presentation of VLPs by antigen presenting cells (APCs). To illustrate the mechanisms, we utilized two-dimensional electrophoresis and tandem MS to compare and identify differentially expressed proteins between hepatitis B virus core antigen VLP (HBc-VLP)-pulsed and control RAW264.7 cells. Of the 25 spots identified as differentially expressed ( p < 0.05) between the two cell lines, 11 (corresponding to 11 unique proteins) were positively identified. Further analysis of two proteins, prohibitin and heat shock protein 70, confirmed that these proteins are expressed at higher levels in HBc-VLP-pulsed RAW264.7 cells compared with control cells. The proteins identified in this study will be useful in revealing the mechanisms that underlie VLP-APC interactions. Overall, this study also provides some useful suggestions for vaccine development and gene therapy.
最近许多研究表明,病毒样颗粒(VLP)在疫苗开发和基因治疗领域有许多潜在应用。然而,我们对抗抗原呈递细胞(APC)呈递VLP所涉及的微妙机制仍知之甚少。为阐明这些机制,我们利用二维电泳和串联质谱来比较和鉴定乙肝病毒核心抗原VLP(HBc-VLP)脉冲处理的RAW264.7细胞与对照RAW264.7细胞之间差异表达的蛋白质。在鉴定出的两株细胞系之间差异表达(p<0.05)的25个斑点中,有11个(对应11种独特蛋白质)得到了明确鉴定。对两种蛋白质——抗增殖蛋白和热休克蛋白70的进一步分析证实,与对照细胞相比,这些蛋白质在HBc-VLP脉冲处理的RAW264.7细胞中表达水平更高。本研究中鉴定出的蛋白质将有助于揭示VLP与APC相互作用的潜在机制。总体而言,本研究也为疫苗开发和基因治疗提供了一些有用的建议。